CN102757513A - Preparation technology of polygonatum polysaccharide peptide - Google Patents

Abstract

The invention discloses a preparation technology of polygonatum polysaccharide peptide. The polygonatum polysaccharide peptide is prepared from polygonatum by extraction, preliminary purification and separation and purification steps, and has the advantages of high purity, low generate cost, good product quality and three less wastes.

Description

The preparation technology of polygonatum polysaccharide peptide

Technical field

The invention belongs to pharmaceutical field, in particular to a kind of preparation technology of polygonatum polysaccharide peptide.

Background technology

Polysaccharide peptide is a kind of gp, is polysaccharide and combination of proteins body, is to contain proteinic VISOSE, generally contains very a high proportion of polysaccharide, and they are counted as sugared ammonia polysaccharide, have very high molecular weight, are made up of the repeating unit of disaccharide.Polysaccharide peptide has very strong biological activity, preliminary study polysaccharide Toplink enhancing body immunologic function, has antitumor action and antioxygenation, but because the research of polygonatum polysaccharide peptide is scarcely out of swaddling-clothes, so other biological function is also undiscovered.

Anti-aging effects

Discover that polygonatum polysaccharide and polysaccharide peptide mixture can reduce the generation of MDA in the mouse aging cerebral tissue; Thereby the generation of oxyradical in the minimizing brain; Strengthen the ability of removing oxyradical; Improve the oxidation resistant function of body, suppress the peroxo-process of body, tissue, cell, and can obviously improve brain cell Na ⁺-K ⁺-ATP enzyme and Ca ²⁺The activity of-ATP enzyme prevents Ca in the cell ²⁺Thereby antidotal effect is played in overload.

Antitumor action

Discover that polysaccharide peptide and radiotherapy merge application, can alleviate radiotherapy side effect, the annual rate of depositing behind the raising advanced esophageal cancer patient treatment; Oral or abdominal injection polysaccharide peptide has restraining effect to mouse Sarcoma-180 ascitic type tumour.Another vitro culture lymphocyte method confirms: polysaccharide peptide can promote the cell proliferation of mice spleen beautiful jade crust, also can strengthen the phagocytic function of mouse macrophage.The antitumor action of prompting polysaccharide peptide and enhancing body cell and humoral immunization function are closely related.Also have research to show: cyclic phosphoric acid amine all has the obvious suppression effect to rat humoral immunization and cellular immunization, and polysaccharide peptide can prevent partly that thymus gland and spleen weight alleviate due to the stop ring phosphorus phthalein amine, and can reduce the decline spoke degree of leukocyte count.

Antioxygenation

Discover; Polysaccharide peptide has anti-oxidant activity preferably, and superoxide anion and hydroxyl radical free radical are had tangible scavenging(action), in the total antioxidation experiment, also shows stronger activity; And be tangible dose-effect relationship; Along with the increase of concentration, scavenging(action) strengthens, but TAC is lower than Vc.

Now, polysaccharide peptide has wide biological activity with it and gets most of the attention, and has attracted a lot of research workers to put into the research of this respect.Especially in healthcare products exploitation and drug development field; The present in this respect research for polysaccharide peptide concentrates on antitumor mostly and improves the immunizing power aspect; But also there is a lot of difficulties as the very big biologically active substance of a kind of molecular weight in polysaccharide peptide in the research aspect the pharmacology, so people temporarily also develop as a kind of healthcare products for the utilization of polysaccharide peptide.But the biological activity that polysaccharide peptide is good will inevitably cause people's attention once more, thereby is the further development and use creation advantageous conditions of polysaccharide peptide.

Summary of the invention

Technical problem to be solved by this invention is to provide a kind of preparation technology of polygonatum polysaccharide peptide.Technical scheme of the present invention is following:

The present invention relates to a kind of preparation technology of polygonatum polysaccharide peptide, it is characterized in that comprising the steps:

1) prepare:

Get the sealwort rhizome and clean, oven dry is preferably dried to constant weight, pulverizes, and the water that adds 10 times of sealwort quality is as extracting solvent;

2) extraction and preliminary purification:

The mixture heating up of sealwort and water to 75-85 ℃, is kept extracting the temperature lixiviate, extract 1-3h at every turn, extract 1-3 time; United extraction liquid, concentrating under reduced pressure TV add H to 1/3-2/3 ₂O ₂Make H ₂O ₂Ultimate density is 0.8-1.2%, and 55-65 ℃ decoloured 20-40 minute, then adds and is equivalent to H ₂O ₂The Na of molar weight 30%-60% ₂SO ₃The trichoroacetic acid(TCA) that adds 0.8-1.2 times of volume 5-7% again shakes up hold over night, 3000-5000rmin ^-1Centrifugal 5-15min gets supernatant, uses molecular weight cut-off to dialyse 12-36 hour as the dialysis tubing of 12000-16000Da, is concentrated into the 10-30% volume then, and it is 60-85%% that adding ethanol makes its alcohol concn, alcohol precipitation, 3000-5000rmin then ^-1Centrifugal 5-15min abandons supernatant, and deposition is the polygonatum polysaccharide peptide extract.

In a preferred implementation of the present invention, described preparation technology also comprises the purification procedures of polysaccharide peptide:

Getting above-mentioned polygonatum polysaccharide peptide extract preparation polysaccharide concentration is the solution of 5-15%; Press polysaccharide peptide: resin quality is than (1: 20-40) go up the strongly basic anion exchange column; Till being eluted to elutriant and adding the nondiscoloration of sulfuric acid anthrone reagent with vaal water then, use 2-4 column volume 0.3-0.5molL again ^-1The NaCl eluant solution, concentrating under reduced pressure elutriant and vacuum lyophilization are the polygonatum polysaccharide peptide.

The present invention is raw material with the sealwort, and through extracting, slightly carrying and purification procedures, resulting polygonatum polysaccharide peptide purity is high, and manufacturing cost is low, good product quality, and the three wastes are few.

Embodiment

Embodiment 1:

1) prepare:

Get the sealwort rhizome and clean, dry to constant weight for 80 ℃, pulverized 60 mesh sieves.Take by weighing a certain amount of sealwort dry powder in the Erlenmeyer flask of refluxing extraction device, the water that adds 10 times of sealwort quality again is as extracting solvent.

2) extraction and preliminary purification:

Erlenmeyer flask moved on to be heated to 80 ℃ on the electric furnace, move to then on the thermostat water bath, keep extracting 80 ℃ of hot water lixiviates of temperature 2h, extract 2 times; United extraction liquid, concentrating under reduced pressure TV to 1/2 adds H ₂O ₂Make H ₂O ₂Ultimate density is 1% (mass percent), and 60 ℃ of constant temperature decoloured 30 minutes, then adds and is equivalent to 1/2 H ₂O ₂The Na of amount (molar weight) ₂SO ₃To remove ydrogen peroxide 50; The trichoroacetic acid(TCA) that adds equal-volume 6% again shakes up hold over night, 4 000rmin ^-1Centrifugal 10min gets supernatant.Use molecular weight cut-off to dialyse 24 hours as the dialysis tubing of 14000Da, be concentrated into 1/5 volume then, it is 80% that adding alcohol makes its ultimate density, and alcohol precipitation spends the night.4 000rmin then ^-1Centrifugal 10min abandons supernatant, and deposition is the polygonatum polysaccharide peptide extract.

3) separation and purification of polysaccharide peptide:

Get a certain amount of polygonatum polysaccharide peptide extract preparation polysaccharide concentration and be 10% solution, press mass ratio (polysaccharide peptide: strongly basic anion exchange column on resin) 1: 30.Till being eluted to elutriant and adding the nondiscoloration of sulfuric acid anthrone reagent with vaal water then, use 3 column volume 0.4molL again ^-1The NaCl wash-out, concentrating under reduced pressure elutriant and vacuum lyophilization are the polygonatum polysaccharide peptide.Detect polysaccharide and protein contnt respectively through sulfuric acid anthrone method and Xylene Brilliant Cyanine G method, wherein protein contnt is about 10%, and polysaccharide content is about 60%, and polygonatum polysaccharide peptide purity can reach more than 70%.

When being understood that; Specific embodiment of the present invention only is a property illustrative purposes presented for purpose of illustration; It limits protection scope of the present invention never in any form; Those skilled in the art can improve or conversion according to above-mentioned explanation, and all these improvement and conversion all should belong to the protection domain of accompanying claims of the present invention.

Claims (2)

1. the preparation technology of a polygonatum polysaccharide peptide is characterized in that comprising the steps:

1) prepare:

Get the sealwort rhizome and clean, oven dry is preferably dried to constant weight, pulverizes, and the water that adds 10 times of sealwort quality is as extracting solvent;

2) extraction and preliminary purification:

The mixture heating up of sealwort and water to 75-85 ℃, is kept extracting the temperature lixiviate, extract 1-3h at every turn, extract 1-3 time; United extraction liquid, concentrating under reduced pressure TV add H to 1/3-2/3 ₂O ₂Make H ₂O ₂Ultimate density is 0.8-1.2%, and 55-65 ℃ decoloured 20-40 minute, then adds and is equivalent to H ₂O ₂The Na of molar weight 30%-60% ₂SO ₃The trichoroacetic acid(TCA) that adds 0.8-1.2 times of volume 5-7% again shakes up hold over night, 3000-5000rmin ^-1Centrifugal 5-15min gets supernatant, uses molecular weight cut-off to dialyse 12-36 hour as the dialysis tubing of 12000-16000Da, is concentrated into the 10-30% volume then, and it is 60-85%% that adding ethanol makes its alcohol concn, alcohol precipitation, 3000-5000rmin then ^-1Centrifugal 5-15min abandons supernatant, and deposition is the polygonatum polysaccharide peptide extract.

2. preparation technology according to claim 1, described preparation technology also comprises the purification procedures of polysaccharide peptide:

Getting above-mentioned polygonatum polysaccharide peptide extract preparation polysaccharide concentration is the solution of 5-15%; Press polysaccharide peptide: resin quality is than (1: 20-40) go up the strongly basic anion exchange column; Till being eluted to elutriant and adding the nondiscoloration of sulfuric acid anthrone reagent with vaal water then, use 2-4 column volume 0.3-0.5molL again ^-1The NaCl eluant solution, concentrating under reduced pressure elutriant and vacuum lyophilization are the polygonatum polysaccharide peptide.